还贝止咳方不同极性部位止咳作用的谱效关系
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摘要
目的:研究还贝止咳方不同极性部位的指纹图谱与其止咳作用的关联性,明确对其药效作用贡献较大的色谱峰,并对色谱峰进行归属。方法:采用梯度洗脱法建立还贝止咳方不同极性部位的色谱峰,以水提物图谱为参照,经色谱工作站处理,得到各部位指纹图谱的共有峰和相应的峰面积。以小鼠咳嗽次数、咳嗽潜伏期及肺灌洗液中的白细胞介素(IL)-4,γ-干扰素(IFN-γ),免疫球蛋白(Ig) E,IL-5的含量为检测指标,对还贝止咳方不同极性部位的止咳作用进行考察。将咳嗽次数、咳嗽潜伏期与共有峰峰面积进行关联性分析,指出对药效作用贡献较大的峰,并根据单味药指纹图谱进行归属。结果:双变量相关分析得出7个对止咳作用贡献较大的色谱峰,分别为12,13,29,32,33,35,39号峰,这些峰所代表的化合物含量越高,止咳作用越强,其中32号峰与咳嗽潜伏期有极显著相关性,13,32号峰与咳嗽次数具有极显著相关性。对这些色谱峰进行归属,发现这些色谱峰分别属于黄芩、还阳参、枳壳、甘草等药味。结论:13号,32号等色谱峰对还贝止咳方的止咳作用发挥的贡献较大,还阳参、黄芩、枳壳对于全方具有显著意义。
Objective: To study the correlation between the fingerprint of different polar parts of Huanbei Zhike recipe and its antitussive effect, identify the chromatographic peaks greatly contributing to its pharmacological effect,and ascribe the chromatographic peaks. Method: Gradient elution method was used to establish the chromatographic peaks of different polar parts of Huanbei Zhike recipe. The common peaks and corresponding peak areas of each part were obtained by referring to the water extract chromatogram and after being processed by chromatographic workstation. The cough frequency,the latent period of cough and the contents of interleukin( IL)-4,interferon-γ( IFN-γ),immunoglobulin( Ig) E and IL-5 in lung lavage fluid of mice were used as the detection indexes to investigate the antitussive effect of different polar parts of Huanbei Zhike recipe.The correlation analysis among cough frequency,cough latent period and common peak area was carried out,and the peaks greatly contributing to the effect of medicine were pointed out,which were assigned according to the fingerprint of single drug. Result: Bivariate correlation analysis showed that seven chromatographic peaks,i. e.No. 12,13,29,32,33,35 and 39,contributed significantly to the antitussive effect. The higher the content of the compounds represented by these peaks,the stronger the antitussive effect. There was a significant correlation between No. 32 peak and the latent period of cough,while No. 13 and No. 32 peaks were significantly correlated with cough frequency. These chromatographic peaks were assigned and found to be present in Scutellaria baicalensis,Crepis Herba,Aurantii Fructus,Glycyrrhiza uralensis,etc. Conclusion: The chromatographic peaks of No. 13 and No. 32 peaks contribute greatly to the antitussive effect of Huanbei Zhike recipe,while Crepis turczaniowii,S. baicalensis and Aurantii Fructus are of significant significance for the whole recipe.
Objective: To study the correlation between the fingerprint of different polar parts of Huanbei Zhike recipe and its antitussive effect, identify the chromatographic peaks greatly contributing to its pharmacological effect,and ascribe the chromatographic peaks. Method: Gradient elution method was used to establish the chromatographic peaks of different polar parts of Huanbei Zhike recipe. The common peaks and corresponding peak areas of each part were obtained by referring to the water extract chromatogram and after being processed by chromatographic workstation. The cough frequency,the latent period of cough and the contents of interleukin( IL)-4,interferon-γ( IFN-γ),immunoglobulin( Ig) E and IL-5 in lung lavage fluid of mice were used as the detection indexes to investigate the antitussive effect of different polar parts of Huanbei Zhike recipe.The correlation analysis among cough frequency,cough latent period and common peak area was carried out,and the peaks greatly contributing to the effect of medicine were pointed out,which were assigned according to the fingerprint of single drug. Result: Bivariate correlation analysis showed that seven chromatographic peaks,i. e.No. 12,13,29,32,33,35 and 39,contributed significantly to the antitussive effect. The higher the content of the compounds represented by these peaks,the stronger the antitussive effect. There was a significant correlation between No. 32 peak and the latent period of cough,while No. 13 and No. 32 peaks were significantly correlated with cough frequency. These chromatographic peaks were assigned and found to be present in Scutellaria baicalensis,Crepis Herba,Aurantii Fructus,Glycyrrhiza uralensis,etc. Conclusion: The chromatographic peaks of No. 13 and No. 32 peaks contribute greatly to the antitussive effect of Huanbei Zhike recipe,while Crepis turczaniowii,S. baicalensis and Aurantii Fructus are of significant significance for the whole recipe.
引文
[1]彭照琪.还阳参乙酸乙酯部位化学成分及HD2部位谱效关联研究[D].太原:山西省中医药研究院,2014.
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[9]石召华,黄文芳,陈立军,等.不同干燥方式对地龙提取物中17种氨基酸的影响[J].中成药,2015,37(5):1135-1138.
[10]佟晓琳,聂颖兰,马琰岩,等.平贝母水煎液化学成分的LC-MSn分析[J].中国实验方剂学杂志,2016,22(21):45-49.
[11]程斌,徐文伟,俞松林,等.浙贝母化痰效应成分指数的建立与应用[J].中国实验方剂学杂志,2017,23(20):71-76.
[12]彭锐,谭均,马鹏,等.太白贝母生物碱的HPLC指纹图谱研究[J].世界科学技术—中医药现代化,2015,17(1):152-155.
[2]彭照琪,岳永花,李小贝,等.还阳参醇提物止咳祛痰平喘及抗炎作用的实验研究[J].药物评价研究,2012,35(6):423-426.
[3]李媛,张雯霞,何盼,等.基于~1H-NMR代谢组学的还贝止咳方对咳嗽变异性哮喘豚鼠的干预作用[J].中草药,2018,49(10):2230-2239.
[4]李媛,张雯霞,刘聪,等.还贝止咳方对肺炎链球菌致大鼠肺热的治疗作用[J].药物评价研究,2018,41(5):788-793.
[5]蔡靓,张倩,杨丰庆.中药谱效学的应用进展[J].中草药,2017,48(23):5005-5011.
[6]刘英,徐永莉,李力.中药谱效关系研究思路与应用概况[J].时珍国医国药,2018,29(3):677-680.
[7]曾令军,林兵,宋洪涛.中药谱效关系研究进展及关键问题探讨[J].中国中药杂志,2015,40(8):1425-1432.
[8]赵清,徐月清,冯天铸,等.不同炮制方法对僵蚕指标性成分的含量影响研究[J].时珍国医国药,2011,22(3):657-660.
[9]石召华,黄文芳,陈立军,等.不同干燥方式对地龙提取物中17种氨基酸的影响[J].中成药,2015,37(5):1135-1138.
[10]佟晓琳,聂颖兰,马琰岩,等.平贝母水煎液化学成分的LC-MSn分析[J].中国实验方剂学杂志,2016,22(21):45-49.
[11]程斌,徐文伟,俞松林,等.浙贝母化痰效应成分指数的建立与应用[J].中国实验方剂学杂志,2017,23(20):71-76.
[12]彭锐,谭均,马鹏,等.太白贝母生物碱的HPLC指纹图谱研究[J].世界科学技术—中医药现代化,2015,17(1):152-155.